J Korean Neurosurg Soc.  2018 Sep;61(5):548-558. 10.3340/jkns.2017.0200.

S100ß, Matrix Metalloproteinase-9, D-dimer, and Heat Shock Protein 70 Are Serologic Biomarkers of Acute Cerebral Infarction in a Mouse Model of Transient MCA Occlusion

Affiliations
  • 1Department of Neurosurgery, Hallym University Kangnam Sacred Heart Hospital, Hallym University College of Medicine, Seoul, Korea.
  • 2Department of Neurosurgery, Korea University Medical Center, Seoul, Korea. sungkha@yahoo.com

Abstract


OBJECTIVE
Diagnosing acute cerebral infarction is crucial in determining prognosis of stroke patients. Although many serologic tests for prompt diagnosis are available, the clinical application of serologic tests is currently limited. We investigated whether S100β, matrix metalloproteinase-9 (MMP-9), D-dimer, and heat shock protein 70 (HSP70) can be used as biomarkers for acute cerebral infarction.
METHODS
Focal cerebral ischemia was induced using the modified intraluminal filament technique. Mice were randomly assigned to 30-minute occlusion (n=10), 60-minute occlusion (n=10), or sham (n=5) groups. Four hours later, neurological deficits were evaluated and blood samples were obtained. Infarction volumes were calculated and plasma S100β, MMP-9, D-dimer, and HSP70 levels were measured using enzyme-linked immunosorbent assay.
RESULTS
The average infarction volume was 12.32±2.31 mm³ and 46.9±7.43 mm³ in the 30- and 60-minute groups, respectively. The mean neurological score in the two ischemic groups was 1.6±0.55 and 3.2±0.70, respectively. S100β, MMP-9, and HSP70 expressions significantly increased after 4 hours of ischemia (p=0.001). Furthermore, S100β and MMP-9 expressions correlated with infarction volumes (p < 0.001) and neurological deficits (p < 0.001). There was no significant difference in D-dimer expression between groups (p=0.843). The area under the receiver operating characteristic curve (AUC) showed high sensitivity and specificity for MMP-9, HSP70 (AUC=1), and S100β (AUC=0.98).
CONCLUSION
S100β, MMP-9, and HSP70 can complement current diagnostic tools to assess cerebral infarction, suggesting their use as potential biomarkers for acute cerebral infarction.

Keyword

Acute ischemic stroke; Biomarkers; Cerebral infarction volume

MeSH Terms

Animals
Biomarkers*
Brain Ischemia
Cerebral Infarction*
Complement System Proteins
Diagnosis
Enzyme-Linked Immunosorbent Assay
Heat-Shock Proteins*
Hot Temperature*
HSP70 Heat-Shock Proteins*
Humans
Infarction
Ischemia
Matrix Metalloproteinase 9*
Mice*
Plasma
Prognosis
ROC Curve
Sensitivity and Specificity
Serologic Tests
Stroke
Biomarkers
Complement System Proteins
HSP70 Heat-Shock Proteins
Heat-Shock Proteins
Matrix Metalloproteinase 9

Figure

  • Fig. 1. A : coronal brain sections. Infarcts are visualized as unstained regions following 2, 3, 5-triphenyltetrazolium chloride staining. No infarct is seen in sham-operated mice. Infarcts are confined to the basal ganglia in 30-minute McAO mice, and extend from the basal ganglia to the cerebral cortex in the 60-minute McAO mice. b : comparison of the infarction volumes between the sham-operated, 30-minute McAO, and 60-minute McAO groups. The infarction volume is significantly larger in the 60-minute McAO group when compared with the 30-minute group (p=0.008). Data are expressed as mean±standard deviation. *p<0.05. McAO : middle cerebral artery occlusion.

  • Fig. 2. comparison of the neurological scores between the shamoperated, 30-minute McAO, and 60-minute McAO groups. Shamoperated mice do not show any neurological deficit. The neurological score is significantly higher in the 60-minute group when compared with the 30-minute group (p=0.032). Data are expressed as mean±standard deviation. *p<0.05. McAO : middle cerebral artery occlusion.

  • Fig. 3. Plasma S100β concentration in all groups. The concentration of S100β increases with occlusion time (p=0.001; A) and correlates significantly with infarct volume (r=0.867, p<0.001; b) and neurological deficit (r=0.802, p<0.001; c). *p<0.05.

  • Fig. 4. Plasma MMP-9 concentration in all groups. The concentration of MMP-9 increases with occlusion time (p=0.001; A) and correlates significantly with infarct volume (r=0.890, p<0.001; b) and neurological deficit (r=0.879, p<0.001; c). *p<0.05. MMP-9 : matrix metalloproteinase-9.

  • Fig. 5. Plasma D-dimer concentration in all groups. There is no significant difference in D-dimer concentration between groups (p=0.843; A), and no correlation with infarct volume (r=0.255, p=0.218; b) or neurological score (r=0.139, p=0.507; c). McAO : middle cerebral artery occlusion.

  • Fig. 6. Plasma HSP70 concentration in all groups. HSP70 concentration increases significantly in both infarction groups (p=0.001) when compared to that of the sham group (A); however, there is no correlation between HSP70 concentration and infarct area (r=0.174, p=0.406; b) or neurological score (r=0.138, p=0.510; c). *p<0.05. HSP70 : heat shock protein 70, McAO : middle cerebral artery occlusion.

  • Fig. 7. Sensitivity and specificity of the biomarkers. MMP-9 and HSP70 both have an AUc of 1. S100β has an AUc of 0.98. The AUc for D-dimer is 0.58. MMP-9 : matrix metalloproteinase-9, HSP70 : heat shock protein 70, AUc : the area under the receiver operating characteristic curve.


Reference

References

1. Ageno W, Finazzi S, Steidl L, Biotti MG, Mera V, Melzi D'Eril G, et al. Plasma measurement of D-dimer levels for the early diagnosis of ischemic stroke subtypes. Arch Intern Med. 162:2589–2593. 2002.
Article
2. Arenillas JF, Alvarez-Sabin J, Molina CA, Chacón P, Fernández-Cadenas I, Ribó M, et al. Progression of symptomatic intracranial large artery atherosclerosis is associated with a proinflammatory state and impaired fibrinolysis. Stroke. 39:1456–1463. 2008.
Article
3. Bader MK, Palmer S. What's the "hyper" in hyperacute stroke? Strategies to improve outcomes in ischemic stroke patients presenting within 6 hours. AACN Adv Crit Care. 17:194–214. 2006.
4. Castellanos M, Leira R, Serena J, Blanco M, Pedraza S, Castillo J, et al. Plasma cellular-fibronectin concentration predicts hemorrhagic transformation after thrombolytic therapy in acute ischemic stroke. Stroke. 35:1671–1676. 2004.
Article
5. Castillo J, Rodriguez I. Biochemical changes and inflammatory response as markers for brain ischaemia: molecular markers of diagnostic utility and prognosis in human clinical practice. Cerebrovasc Dis 17 Suppl. 1:7–18. 2004.
Article
6. Foerch C, Otto B, Singer OC, Neumann-Haefelin T, Yan B, Berkefeld J, et al. Serum S100B predicts a malignant course of infarction in patients with acute middle cerebral artery occlusion. Stroke. 35:2160–2164. 2004.
Article
7. Foerch C, Singer OC, Neumann-Haefelin T, du Mesnil de Rochemont R, Steinmetz H, Sitzer M. Evaluation of serum S100B as a surrogate marker for long-term outcome and infarct volume in acute middle cerebral artery infarction. Arch Neurol. 62:1130–1134. 2005.
Article
8. Fujii Y, Takeuchi S, Harada A, Abe H, Sasaki O, Tanaka R. Hemostatic activation in spontaneous intracerebral hemorrhage. Stroke. 32:883–890. 2001.
Article
9. Geiger S, Holdenrieder S, Stieber P, Hamann GF, Bruening R, Ma J, et al. Nucleosomes as a new prognostic marker in early cerebral stroke. J Neurol. 254:617–623. 2007.
Article
10. Hacke W, Kaste M, Bluhmki E, Brozman M, Dávalos A, Guidetti D, et al. Thrombolysis with alteplase 3 to 4.5 hours after acute ischemic stroke. N Engl J Med. 359:1317–1329. 2008.
Article
11. Jiang SX, Lertvorachon J, Hou ST, Konishi Y, Webster J, Mealing G, et al. Chlortetracycline and demeclocycline inhibit calpains and protect mouse neurons against glutamate toxicity and cerebral ischemia. J Biol Chem. 280:33811–33818. 2005.
Article
12. Jin X, Xiao C, Tanguav RM, Yang L, Wang F, Chen M, et al. Correlation of lymphocyte heat shock protein 70 levels with neurologic deficits in elderly patients with cerebral infarction. Am J Med. 117:406–411. 2004.
Article
13. Joly AL, Wettstein G, Mignot G, Ghiringhelli F, Garrido C. Dual role of heat shock proteins as regulators of apoptosis and innate immunity. J Innate Immun. 2:238–247. 2010.
Article
14. Kleindienst A, Ross Bullock M. A critical analysis of the role of the neurotrophic protein S100B in acute brain injury. J Neurotrauma. 23:1185–1200. 2006.
Article
15. Laskowitz DT, Kasner SE, Saver J, Remmel KS, Jauch EC, BRAIN Study Group. Clinical usefulness of a biomarker-based diagnostic test for acute stroke: the Biomarker Rapid Assessment in Ischemic Injury (BRAIN) study. Stroke. 40:77–85. 2009.
Article
16. Lynch JR, Blessing R, White WD, Grocott HP, Newman MF, Laskowitz DT. Novel diagnostic test for acute stroke. Stroke. 35:57–63. 2004.
Article
17. Maas MB, Furie KL. Molecular biomarkers in stroke diagnosis and prognosis. Biomark Med. 3:363–383. 2009.
Article
18. Rajdev S, Hara K, Kokubo Y, Mestril R, Dillmann W, Weinstein PR, et al. Mice overexpressing rat heat shock protein 70 are protected against cerebral infarction. Ann Neurol. 47:782–791. 2000.
Article
19. Reynolds MA, Kirchick HJ, Dahlen JR, Anderberg JM, McPherson PH, Nakamura KK, et al. Early biomarkers of stroke. Clin Chem. 49:1733–1739. 2003.
Article
20. Rosell A, Ortega-Aznar A, Alvarez-Sabín J, Fernández-Cadenas I, Ribó M, Molina CA, et al. Increased brain expression of matrix metalloproteinase-9 after ischemic and hemorrhagic human stroke. Stroke. 37:1399–1406. 2006.
Article
21. Schmidt-Kastner R, Zhang B, Belayev L, Khoutorova L, Amin R, Busto R, et al. DNA microarray analysis of cortical gene expression during early recirculation after focal brain ischemia in rat. Brain Res Mol Brain Res. 108:81–93. 2002.
Article
22. Skoloudik D, Bar M, Sanák D, Bardon P, Roubec M, Langová K, et al. D-dimers increase in acute ischemic stroke patients with the large artery occlusion, but do not depend on the time of artery recanalization. J Thromb Thrombolysis. 29:477–482. 2010.
Article
23. Snider BJ, Du C, Wei L, Choi DW. Cycloheximide reduces infarct volume when administered up to 6 h after mild focal ischemia in rats. Brain Res. 917:147–157. 2001.
Article
24. Swanson RA, Morton MT, Tsao-Wu G, Savalos RA, Davidson C, Sharp FR. A semiautomated method for measuring brain infarct volume. J Cereb Blood Flow Metab. 10:290–293. 1990.
Article
25. Whiteley W, Chong WL, Sengupta A, Sandercock P. Blood markers for the prognosis of ischemic stroke: a systematic review. Stroke. 40:e380–e389. 2009.
26. Whiteley W, Wardlaw J, Dennis M, Lowe G, Rumley A, Sattar N, et al. Blood biomarkers for the diagnosis of acute cerebrovascular diseases: a prospective cohort study. Cerebrovasc Dis. 32:141–147. 2011.
Article
27. Wunderlich MT, Wallesch CW, Goertler M. Release of neurobiochemical markers of brain damage is related to the neurovascular status on admission and the site of arterial occlusion in acute ischemic stroke. J Neurol Sci. 227:49–53. 2004.
Article
28. Yenari MA, Giffard RG, Sapolsky RM, Steinberg GK. The neuroprotective potential of heat shock protein 70 (HSP70). Mol Med Today. 5:525–531. 1999.
Article
29. Zhan X, Ander BP, Liao IH, Hansen JE, Kim C, Clements D, et al. Recombinant Fv-Hsp70 protein mediates neuroprotection after focal cerebral ischemia in rats. Stroke. 41:538–543. 2010.
Article
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